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Bacterial Susceptibility to Antibiotics Joseph P Henry Ford Hospital Medical Journal Volume 6 | Number 2 Article 6 6-1958 Bacterial Susceptibility To Antibiotics Joseph P. Truant Follow this and additional works at: https://scholarlycommons.henryford.com/hfhmedjournal Part of the Life Sciences Commons, Medical Specialties Commons, and the Public Health Commons Recommended Citation Truant, Joseph P. (1958) "Bacterial Susceptibility To Antibiotics," Henry Ford Hospital Medical Bulletin : Vol. 6 : No. 2 , 189-195. Available at: https://scholarlycommons.henryford.com/hfhmedjournal/vol6/iss2/6 This Article is brought to you for free and open access by Henry Ford Health System Scholarly Commons. It has been accepted for inclusion in Henry Ford Hospital Medical Journal by an authorized editor of Henry Ford Health System Scholarly Commons. For more information, please contact [email protected]. BACTERIAL SUSCEPTIBILITY TO ANTIBIOTICS JOSEPH P. TRUANT* The determination of in vitro susceptibility testing of bacteria to antibiotics has been of considerable value in assisting the clinician to choose the most effective thera­ peutic agent in order to overcome an infectious process. A great deal of valuable time may be lost, often to the detriment of the patient if bacterial cultures and in vitro susceptibility tests are not initiated as soon as possible. The information gained by following this procedure is not only important as an aid in determining which anti­ biotic may be effective against a possible pathogen, but also which antibiotics are in­ effective. As the scope of antibiotic therapy has been expanded by the discovery and de­ velopment of many new chemotherapeutic agents, the problem of selecting the most suitable one for any individual case has become more and more complex. As a result, the following system has been adopted in the bacteriology laboratory. Routinely, with requests for sensitivity, the pathogenic bacteria are tested in one, two or possibly three stages. If stage one shows the organism to be sensitive to one of the antibiotics tried, stage two will not be undertaken unless there is a specific request by the clinician. For Gram-positive bacteria, the first stage includes testing with sensitivity discs which contain penicillin, dihydrostreptomycin, tetracycline and chloramphenicol. In the second stage erythromycin, novobiocin and occasionally oleandomycin (matromy- cin) are employed. In reserve are such anti-bacterial agents as bacitracin, furacin, furadantin, neomycin and the sulfas which are employed only after consultation with the clinician. For the Gram-negative bacteria the first stage includes dihydrostreptomycin, tetracycline and chloramphenicol. Strains of Proteus are tested wflh penicillin and Pseudomonas strains are tested with polymyxin in addition to the mentioned antibiotics during the first trial. In the second stage the clinician may choose from the following; furacin, furadantin, neomycin, polymyxin and the sulfas. The sensitivity of any given strain or organism to an antimicrobial agent is generally defined in terms of concentration of the agent which inhibits the growth partially or completely. Several methods of determining this value are in common use and the results obtained by the different methods may show some discrepancy, depending on the details of the method and the choice of end point. In this laboratory, we employ the disc technique since it provides a satisfactory and practical routine procedure for the sensitivity evaluation of microorganisms to a variety of chemotherapeutic agents. The antibiotic discs most commonly employed are listed in Table I. It should be emphasized that this technique is a qualitative pro­ cedure and is designed to determine whether the organisms are sensitive or resistant according to the following scheme; "Division of Microbiology, Department of Laboratories 189 Bacterial Susceptibility to Antibiotics Sensitive — a marked zone of inhibition around the discs of both high and low series. Moderately sensitive — minimal if any detectable zone of inhibition around the low series disc and a distinct zone of inhibition around the high series disc. Resistant — absence of a zone of inhibition around the discs of both con­ centrations. TABLE I CONCENTRATIONS OF ANTIBIOTIC DISCS Low series High Series Penicillin 2 units 10 units Dihydrostreptomycin 10 meg. 50 meg. Tetracycline 5 meg. 30 meg. Chloramphenicol 5 meg. 30 meg. Erythromycin 2 meg. 15 meg. Novobiocin 5 meg. 30 meg. Oleandomycin 2 meg. 15 meg. Neomycin 5 meg. 30 meg. Polymyxin 5 meg. 30 meg. Furacin 100 meg. 100 meg. Furadantin 100 100 n:c-. During the past year our interest has been centered on methods for controlling the variables inherent in routine disc sensitivity testing. A large number of Gram- positive and Gram-negative isolates from the routine services were tested for sensi­ tivity to several antibiotics by three different methods. In this manner it was possible to reveal the frequency, extent and direction of the variations in the results obtained with the different procedures as applied to antibiotics which are now in general use. It was hoped, in this manner also to elucidate some of the factors responsible for the discrepancies which occur occasionafly. MATERIALS AND METHODS Two hundred and thirty bacterial strains and ten antibacterial agents were included in this study. The three methods which were employed for susceptibility testing were as follows; (1) The filter paper "disc agar diffusion" technique (2) The tube or broth dilution technique (3) The cylinder-plate method The details concerning these procedures are clearly discussed by Grove and Randall (1) as well as by Jackson et al (2). 190 Truant In order to obtain the best comparison and evaluation of the different methods, the tube dflution and the cylinder-plate tests done with any given combination of organisms and antibiotics, were set up and carried out using the same seed culture and the same freshly prepared dflutions of antibiotics. From three to six organisms were tested with the antibiotics at one time. The tests employing the medicated discs were carried out separately in our laboratory and some .strains were also tested by the Division of Labora­ tories, Michigan Department of Health. The results were also compared with those reported by the technicians on the routine services. RESULTS It is not feasible to record the results of all tests with every combination of organisms and antibiotics included in this study. However, sufficient data will be reported in tabular form to illustrate the highlights which were brought out by this investigation. Table II shows the results of testing 130 staphylococci against penicillin, tetra­ cycline, dihydrostreptomycin. chloramphenicol, oleandomycin, erythromycin and novo­ biocin by the three methods. Inspection of the data shows only small differences in the values obtained by the different procedures. Thus, in almost all instances, agreement was observed. Minor discrepancies (one-dilution differences) occasionally would have altered the reporting in lieu of changing the sensitivity pattern from one category to another. This is practically only important with organisms on the borderline between moderately resistant and resistant, since the latter interpretation would exclude therapy. The antibiotic spectrum of the staphylococci shown in Table II may be of practical value since it is based on a large number of recent isolates obtained from specimens collected in this hospital. From this spectrum it can be seen that as many as 78.8 per cent of the isolates may be resistant to penicillin. The spectrums of the other anti­ biotics shown in Table II correspond to tho.se reported by other investigators. Table III shows the results of the susceptibility tests wflh the Gram-negative bacilli. The data shows a greater variation in the sensitivity pattern due to the different methods than the results obtained using similar procedures against the staphylococci. The greatest discrepancy occurred with dihydrostreptomycin using the tube dilution procedure. According to Garrod (4) and others, the levels of resistance to streptomycin with in vitro susceptibility tests may vary with changes in the size of the inoculum. Branch et al (5) have shown that the size of the inoculum is an important factor in changing the minimum inhibitory concentration in the tube dilution method and it also has a slight effect in the agar well method but has not been found to exert any demonstrable changes in the reading of the 'disc results'. Furthermore, as already noted by others (2, 3). additional factors such as media, period of incubation, choice of size of inhibition zone etc. may account for some of the variation in the results shown in Table IIL The data in Table III shows dihydrostreptomycin and furacin were the most effective antibiotics in vitro against the Gram-negative bacilli. The least effective of the five agents was tetracycline. Previous experience has shown that peniciflin is ineffective against this group of organisms. Occasionafly high concentrations of penicillin will inhibit members of the genus Proteus. 191 TABLE II COMPARATIVE TESTS BY THREE lETHODS ON 130 STAPHYLOCOCCI 5: Penicillin Tetracycline Dihydro- Chloramphenicol Hatromycin Erythromycin Novobiocin streptomycin METHODS DISC ^, % sensitive^,j^^ 21.2 42.4 44.3 90.0 97.7 93.4 100 -: % resistant 78.8 57.6 55.7 10.0 2,3 6.6 0 ••3 TUBE DILUTIOI % sensitive 18.2 42.4 44.3 88.6 97.7 90.4 100 % resistant 81.8 57.6 55.7 11.4 2.3 9.6 0 CYLINDER-PLATE % sensitive 22.7 39.4 44.3 87.1 95.5 93.4 100 % resistant
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